Method to prepare dye-based reference material

Abstract

Provided is a method of preparing dye-based reference materials useful for calibrating or qualifying instrument systems that are diagnostic spectroscopically for hemoglobin and CO-ox fractions. The dye-based reference materials are non-proteinaceous and may be interpreted by CO-oximeter instrument systems as providing a spectrum substantially equivalent to the spectrum of blood, particularly with regard to hemoglobin and hemoglobin CO-ox fractions. Dye-based reference materials may be formulated to provide CO-ox fractions approximating those seen in a clinical setting, including normal physiological CO-ox fractions. The dye-based reference materials may be combined with other ingredients thus making the materials useful as references for pH and blood gas instruments and/or electrolyte instruments in addition to CO-oximeter instrument systems.

That which is claimed is: 
     
       1. A reference material comprising (a) a first dye having a main absorption maxima at a wavelength between about 560 to about 580 nm wherein said first dye is a red dye selected from the group consisting of sulforhodamine B, chlorophenol red, xylenol orange, and sulforhodamine 101; (b) a second dye having an absorption maxima at a wavelength between about 350 to about 450 mn wherein said second dye is a yellow dye selected from the group consisting of mordant yellow 7, tartrazine, orange G, hydroxypyrenetrisulfonic acid, mordant yellow 10, combinations thereof; (c) a third dye having an absorption maxima at a wavelength between about 625 and about 640 nm wherein said third dye is selected from the group consisting of brilliant blue FC (FD&C blue 1), erioglaucine, lissamine, alphazurine A, patent blue violet, patent blue VF, hydroxy naphthol blue, and combination thereof; and (d) a fourth dye having an absorption maxima at a wavelength between about 700 to about 780 nm and an absorption band width of from about 150 to about 200 nm wherein said fourth dye is selected from the group consisting of IR 125, naphthol green B, and combination thereof. 
     
     
       2. A material according to claim 1 wherein said first dye is sulforhodamine B; said second dye is mordant yellow 7; said third dye is patent blue violet; and said fourth dye is naphthol green B. 
     
     
       3. A material according to claim 1 wherein said first dye is sulforhodamine B; said second dye is tartrazine; said third dye is patent blue violet; and said fourth dye is naphthol green B. 
     
     
       4. A material according to claim 1 wherein said first dye is sulforhodamine B; said second dye is mordant yellow 10; and said third dye is patent blue VF; and said fourth dye is naphthol green B. 
     
     
       5. A material according to claim 1 wherein said first dye is sulforhodamine B; said second dye is mordant yellow 7; and said third dye is FD&C blue 1; and said fourth dye is naphthol green B. 
     
     
       6. A material according to claim 1 wherein the first dye is used in an amount ranging from about 0.55 mM to about 4 mM; the second dye is used in an amount ranging from about 1 mM to about 10 mM; the third dye is used in an amount ranging from about 0.01 mM to 0.3 mM; and the fourth dye is used in an amount ranging from 0 to about 2.5 mM. 
     
     
       7. A material according to claim 1 comprising the following:   ______________________________________                                    
COMPOUND      CONCENTRATIONS                                              
______________________________________                                    
NaCl          110 to       160 mM                                         
KCl           2.5 to       7.5 mM                                         
CaCl.sub.2    2 to         4 mM                                           
HEPES         35 to        40 mM                                          
TRIS.HCl      15 to        40 mM                                          
NaOH          20 to        50 mM                                          
EDTA          1 to         1.7 mM                                         
NaHCO.sub.3   15 to        20 mM                                          
LiCl          10 to        25 mM                                          
Glucose       0.5 to       10 g/L                                         
Li lactate    0.5 to       14 mM                                          
Sulforhodamine B                                                          
              0.55 to      1.3 g/L                                        
Mordant Yellow 7                                                          
              1.4 to       3 g/L                                          
Patent Blue Violet                                                        
              0.07 to      0.08 g/L                                       
Naphthol Green B                                                          
              0.4 to       1.1 g/L.                                       
______________________________________                                    
     
     
     
       8. A material according to claim 7 wherein said reference material provides clinical hemoglobin fractions of: about 60% to about 98% oxyhemoglobin; from 0 to about 20% carboxyhemoglobin; from 0 to about 20% methemoglobin; and from 0 to about 20% reduced hemoglobin. 
     
     
       9. A material according to claim 7 wherein said reference material provides physiological hemoglobin fractions of: oxyhemoglobin 94-98%, carboxyhemoglobin 0 about 1%, methemoglobin 0 to about 1.5%, and reduced hemoglobin from about 1 to about 5%. 
     
     
       10. A method for quality control of hemoglobin instrument systems comprising: (1) subjecting said instrument to quality control standard material comprising (a) a first dye having a main absorption maxima at a wavelength between about 560 to about 580 nm; (b) a second dye having an absorption maxima at a wavelength between about 350 to about 450 nm; and (c) a third dye having an absorption maxima at a wavelength between about 625 and about 640 nm; and (d) a fourth dye having an absorption maxima at a wavelength between about 700 to about 780 nm and an absorption band width of from about 150 to about 200 nm; wherein said dyes are used in a sufficient amount to provide said control with a pre-determined level of hemoglobin fractions; (2) obtaining an instrument measurement of hemoglobin fractions of said control; and (3) comparing said instrument measurement off said hemoglobin fractions of said control with said predetermined level of hemoglobin fractions to check the accuracy of said instrument, wherein said quality control standard material is prepared with at least three levels and wherein at least one level has said fourth dye, said three levels of quality control standard material comprising the following:   ______________________________________                                    
Dye            Level a   Level b   Level c                                
______________________________________                                    
Sulforhodamine B                                                          
               0.98 mM   1.95 mM   2.27 mM                                
Mordant Yellow 7                                                          
               6.65      7.63      3.89                                   
Patent Blue Violet                                                        
               0.14      0.11      0.14                                   
Naphthol Green B                                                          
               0         1.10      0.48.                                  
______________________________________                                    
       
     
     
       11. A method for quality control of hemoglobin instrument systems comprising: (1) subjecting said instrument to a quality control standard material comprising: (a) a first dye having a main absorption maxima at a wavelength between about 560 to about 580 nm; (b) a second dye having an absorption maxima at a wavelength between about 350 to about 450 nm; and (c)a third dye having an absorption maxima at a wavelength between about 625 and about 640 nm; and (d) a fourth dye having an absorption maxima at a wavelength between about 700 to about 780 nm and an absorption band width of from about 150 to about 200 nm; wherein said dyes are used in a sufficient amount to provide said control with a pre-determined level of hemoglobin fractions; (2) obtaining an instrument measurement of hemoglobin fractions of said control; and (3) comparing said instrument measurement of said hemoglobin fractions of said control with said predetermined level of hemoglobin fractions to check the accuracy of said instrument, wherein said quality control standard material is prepared with at least three levels and wherein at least one level has said fourth dye, said three levels of quality control standard material comprising the following:   ______________________________________                                    
Dye            Level a   Level b   Level c                                
______________________________________                                    
Sulforhodamine B                                                          
               1.27 mM   2.53 mM   3.22 mM                                
Tartrazine     1.34      1.87      1.60                                   
Patent Blue Violet                                                        
               0.11      0.17      0.25                                   
Naphthol Green B                                                          
               0.98      1.96      0.64.                                  
______________________________________                                    
       
     
     
       12. A method for quality control of hemoglobin instrument systems comprising: (1) subjecting said instrument to a quality control standard material comprising (a) a first dye having a main absorption maxima at a wavelength between about 560 to about 580 nm; (b) a second dye having an absorption maxima at a wavelength between about 350 to about 450 nm; and (c) a third dye having an absorption maxima at a wavelength between about 625 and about 640 nm; and (d) a fourth dye having an absorption maxima at a wavelength between about 700 to about 780 mn and an absorption band width of from about 150 to about 200 nm; wherein said dyes are used in a sufficient amount to provide said control with a pre-determined level of hemoglobin fractions; (2) obtaining an instrument measurement of hemoglobin fractions of said control: and (3) comparing said instrument measurement of said hemoglobin fractions of said control with said predetermined level of hemoglobin fractions to check the accuracy of said instrument, wherein said quality control standard material is prepared with at least three levels and wherein at least one level has said fourth dye said three levels of quality control standard material comprising the following:   ______________________________________                                    
Dye            Level a   Level b   Level c                                
______________________________________                                    
Sulforhodamine B                                                          
               1.17 mM   1.97 mM   2.19 mM                                
Mordant Yellow 7                                                          
               5.33      6.14      2.62                                   
FD&C Blue 1    0.03      0.04      0.03                                   
Naphthol Green B                                                          
               0.97      2.30      0.14.                                  
______________________________________                                    
       
     
     
       13. A method for quality control of hemoglobin instrument systems comprising: (1) subjecting said instrument to a quality control standard material comprising (a) a first dye having a main absorption maxima at a wavelength between about 560 to about 580 nm; (b) a second dye having an absorption maxima at a wavelength between about 350 to about 450 nm; (c) a third dye having an absorption maxima at a wavelength between about 625 to about 640 nm; and a fourth dye having an absorption maxima at a wavelength between about 700 to about 780n m and an absorption band width of from about 150 to about 200 nm; wherein said dyes are used in a sufficient amount to provide said control with a predetermined level of hemoglobin fractions; (2) obtaining an instrument measurement of hemoglobin fractions of said control, and (3) comparing said instrument measurement of said hemoglobin fractions of said control with said predetermined level of hemoglobin fractions to check the accuracy of said instrument, wherein said quality control standard material is prepared with at least three levels and wherein at least one level has said fourth dye, said three levels of quality control standard material comprising the following:-   ______________________________________                                    
Dye            Level a   Level b   Level c                                
______________________________________                                    
Sulforhodamine B                                                          
               1.17 mM   1.97 mM   2.19 mM                                
Mordant Yellow 7                                                          
               5.33      6.14      2.62                                   
FD&C Blue 1    0.03      0.04      0.03                                   
Naphthol Green B                                                          
               0.97      2.30      0.14.                                  
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